Analysis of the expression profiles of Marek's disease virus-encoded microRNAs by real-time quantitative PCR.

MicroRNAs (miRNAs) are a large class of endogenous approximately 22-nucleotide long non-coding RNAs involved in post-transcriptional silencing of gene expression by translational repression or direct cleavage of the target mRNAs. Several hundreds of miRNAs have now been identified in a wide range of organisms including many viruses. Marek's disease virus serotype 1 (MDV-1) is a highly contagious oncogenic herpesvirus that induces fatal T-cell lymphomas in chickens. MDV-1 has been shown recently to encode several miRNAs clustered in the Meq and LAT region of the viral genome. The majority of these miRNAs are expressed at very high levels in infected chicken embryo fibroblasts (CEF) as well as in MDV-transformed lymphomas and lymphoblastoid cell lines derived from tumours. As part of the studies to evaluate the functions of these miRNAs, a quantitative RT-PCR to analyse the expression of three MDV-encoded miRNAs, viz. miR-4, miR-8 and miR-12 in CEF infected with oncogenic strain RB-1B and vaccine strain CVI988 was developed. The study shows that these miRNAs are expressed at very high levels in the tumour tissues compared to the non-tumour tissues from the infected birds indicating that expression levels of MDV-1 miRNAs can be used as biomarkers for transformation.